GSM7926037: AML2_sgLuc_VTP50469_H2AK119ub; Homo sapiens; ChIP-Seq
Briefly, 20-50 million cells were cross-linked by formaldehyde (1%) and then quenched with 0.125 M glycine. After washing with cold PBS, cells were suspended in lysis buffer (50 mM HEPES pH 7.5, 140 mM NaCl, 1 mM EDTA, 10% glycerol, 0.5% NP-40, 0.25% TritonX-100, and protease inhibitor cocktail) and subjected to sonication to shear the chromatin DNA into sizes ranging from 200 to 500 bp. After centrifugation, the supernatant was incubated with primary antibody and Protein A/G Dynabeads magnetic beads (Invitrogen) at 4°C overnight. After six times wash with RIPA wash buffer (50 mM HEPES-KOH, pH 7.6, 500 mM LiCl, 1 mM EDTA, 1% NP-40, and 0.7% Na-Deoxycholate) and one time wash with TE buffer (10 mM Tris-HCl pH 8.0, 1 mM EDTA, and 50 mM NaCl), chromatin was eluted from beads in elution buffer (50 mM Tris-HCl pH 8.0, 10 mM EDTA, 1% SDS). After reverse crosslinking, ChIP DNA was purified using a DNA clean-up kit (Zymo Research). NEBNext Ultra II Library Prep kit was used for library preparation. ChIP libraries were sequenced on the NovaSeq PE150 platform.